Can anyone suggest the number of A549 cells and a procedure for doing in vivo (mice) tumor regression assays?
We normally use 500.000 to 1000000 cells (they should be in the exponential phase, so I wouldn't use cells from flasks with > 80% confluence) and let them form lesions of about 0.8x0.8 cm surface area. These cells (in nude mice) grow well, but not as fast as H460 cells for instance
@ Dear Lorenzo: Thanks...How many days they took to form the tumor and moreover can i do the regression assay with transient transfection (with lipofec...) ???
Hi Paul,
sorry for answering you so late, I rarely connect here and I received any notifications. To answer your questions:
- tumors generally arise 7-12 days upon inoculation (obviously with some variations depending on the general status and precise number of cells at inoculation).
- transient transfection with oligofectamine and lipofectamine works perfectly with these cells in vitro, but be prepared to lose the effects of siRNA/plasmid in the BEST case after 4-5 days (worst case = 1-2 days)
Dear Lorenzo : Thanks for ur help. i haven't done the expt because i am experiencing a strange problem that A549 cells are prone to death upon any transfection,[ Any construct transfection leads to heavy cell death} (I generally used Lipofectamine 2000, .3 million cells in 60 mm ), Can you help me out of this..
Hi Paul, pretty weird, indeed: I've always transfected them with Lipofectamine 2000 without major (>15%) extents of cell death. Normally, I transfected them in 6w plates, at 90% confluence, or so, with 4-8 microL lipofect per well plus 1 microg DNA...
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